Objective: To observe the expression of TK1, Ki-67 and p53 in breast cancer (BC) and their prognosis. Methods: Sixty patients with breast cancer admitted from March 2009 to May 2013 underwent radical mastectomy and their histological specimens were obtained. The expressions of TK1, Ki-67 and p53 in these breast cancer tissues and their adjacent tissues were examined by immunohistochemical staining, and in combination with the follow-up data, the relations of TK1, Ki-67 and p53 expression with the prognosis of the patients were analyzed. Results: The positive expression rates of TK1, Ki-67 and p53 in breast cancer tissues were all significantly higher than those in their adjacent tissues (all P<0.05). The expression intensities of TK1, Ki-67 and p53 markedly affected the 5-year survival rate of the patients, which all indicated that the higher the expression intensity, the lower the survival rate (all P<0.05). The positive expression rates of TK1, Ki-67 and p53 in patients with recurrence or metastasis within 5 years were significantly higher than those in patients without recurrence or metastasis. Conclusion: The expressions of TK1, Ki-67 and p53 are enhanced in breast cancer tissue, and their expression intensities are closely associated with unfavorable prognosis of the patients.